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The Evolution of Immunocytochemistry in the Dissection of Neural Complexity

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Immunocytochemistry and Related Techniques

Part of the book series: Neuromethods ((NM,volume 101))

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Abstract

After more than 70 years from its initial development, immunocytochemistry (ICC ) has become a fundamental technique in the study of the nervous system . After a brief excursus along the history of the different techniques that led to substantial amelioration of the original indirect immunofluorescence protocol, we discuss here the main advantages and disadvantages of the individual techniques for the study of central and peripheral neurons, in parallel with standardization , quantification , and reaction bias . Particular attention is given to immunofluorescence and its novel developments that allow high-resolution imaging at the light microscope level. The possibility of combining ICC with other fundamental techniques for analysis of neuronal circuitry such as neurotracing , electrophysiology , and molecular biology is also discussed, as well as a series of approaches for correlative light and electron microscopic studies.

The emerging picture is that ICC still represents an invaluable tool for histological and cytological analysis of neural complexity.

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Notes

  1. 1.

    The term was coined by Paul Ehrlich at the end of the nineteenth century when he developed in his side-chain theory to explain the immune response-->.

  2. 2.

    Albeit the terms fluorochrome--> and fluorophore are often used as synonyms, we will use fluorochrome to indicate a fluorescent dye or protein used either directly as a specimen stain or conjugated to a biologically active substance to make a fluorophore (fluorescent probe).

  3. 3.

    John William Strutt, third Baron Rayleigh, 1842–1919

  4. 4.

    The existence of the stimulated emission--> process was first theoretically postulated by Einstein back in 1917. Now it is recognized as an universal optical process in which a molecule at its excited state can be stimulated down to its ground state by an incident photon with proper frequency, simultaneously creating a new coherent photon with the same phase, frequency, polarization, and direction as the incident one.

  5. 5.

    An evanescent wave is a near-field standing wave with an intensity that exhibits exponential decay--> with distance from the boundary at which the wave was formed.

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Authors and Affiliations

  1. Department of Veterinary Sciences and National Institute of Neuroscience, University of Turin, Via Leonardo da Vinci 44, 10095, Grugliasco, Turin, Italy

    Adalberto Merighi Ph.D. & Laura Lossi M.D., Ph.D.

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  1. Adalberto Merighi Ph.D.
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  2. Laura Lossi M.D., Ph.D.
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Correspondence to Laura Lossi M.D., Ph.D. .

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  1. Department of Veterinary Sciences, University of Torino, Grugliasco, Torino, Torino, Italy

    Adalberto Merighi

  2. Department of Veterinary Sciences, University of Torino, Grugliasco, Torino, Torino, Italy

    Laura Lossi

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Merighi, A., Lossi, L. (2015). The Evolution of Immunocytochemistry in the Dissection of Neural Complexity. In: Merighi, A., Lossi, L. (eds) Immunocytochemistry and Related Techniques. Neuromethods, vol 101. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2313-7_1

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  • DOI: https://doi.org/10.1007/978-1-4939-2313-7_1

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